芦荟大黄素通过激活Caspase-3/GSDME焦亡通路发挥抗胶质瘤作用  

Aloe-emodin suppresses glioblastoma by activating Caspase-3/GSDME pyroptosis pathway

作　　者：方兴刚 汪嫚[2] 陈卓 张思思 罗杰 FANG Xing-gang;WANG Man;CHEN Zhuo;ZHANG Si-si;LUO Jie(Clinical College of Traditional Chinese Medicine,Hubei University of Chinese Medicine,Wuhan 430065;Hubei Key Laboratory of Embryonic Stem Cells,Taihe Hospital Affiliated to Hubei University of Medicine,Shiyan Hubei 442000;Pharmacy and Nursing Institute,Hubei University of Medicine,Shiyan Hubei 442000)

机构地区：[1]湖北中医药大学中医临床学院,武汉430065 [2]胚胎干细胞研究湖北省重点实验室,湖北医药学附属太和医院,湖北十堰442000 [3]湖北医药学院药护学院,湖北十堰442000

出　　处：《中南药学》2023年第2期291-297,共7页Central South Pharmacy

基　　金：国家自然科学基金面上项目(No.81671831);湖北省2020年引进国外人才和智力项目(No.Yz2020015);2022年度湖北省教育厅科学研究计划指导性项目(No.B2022131)。

摘　　要：目的 探讨芦荟大黄素(AE)抗脑胶质母细胞瘤(GBM)的作用及机制。方法 梯度浓度AE处理体外培养的GBM细胞,MTT实验检查GBM细胞活力,倒置显微镜及LDH释放实验检测AE对GBM细胞的焦亡诱导作用;Western blot实验检测AE对Caspase-3、GSDME蛋白的活化作用,qPCR检测AE对GBM细胞GSDME在m RNA水平的调控作用;DCFH-DA标识GBM细胞活性氧,荧光显微镜及流式细胞术检测AE处理的GBM细胞活性氧产生水平,JC-1试剂盒检测线粒体膜电位。结果 与对照组相比,AE能显著降低GBM细胞活性(P <0.05),诱导GBM细胞出现焦亡特异性外观和LDH释放,剪切活化Caspase-3和GSDME蛋白,在mRNA水平升高GSDME的表达,诱导GBM细胞活性氧产生及线粒体膜电位降低。结论 芦荟大黄素通过激活Caspase-3/GSDME焦亡通路诱导GBM焦亡发挥抗胶质瘤作用。Objective To determine the anti-glioblastoma(GBM) effect and mechanism of aloe-emodin(AE). Methods GBM cells were treated with different concentrations of AE;the cell viability was tested by MTT assay;the pyroptosis-inducing effect was detected by inverted microscopy and lactic acid dehydrogenase(LDH) release assay;the activation of Caspase-3 and GSDME proteins was detected by Western blot;the effect of AE on the regulation of GSDME at mRNA level was detected by qPCR;the level of reactive oxygen species(ROS) in AE treated GBM cells was detected by fluorescence microscopy and flow cytometry by the DCFH-DA markers;and the mitochondrial membrane potential was detected by JC-1 kit. Results Compared with the control group, AE significantly reduced the viability of GBM cells(P < 0.05), induced pyroptosis-specific appearance and lactic acid dehydrogenase release, activated Caspase-3 and GSDME protein, elevated GSDME expression at mRNA levels, increased cellular reactive oxygen species, and decreased mitochondrial membrane potential. Conclusion AE suppresses glioblastoma by activating Caspase-3/GSDME pyroptosis pathway.

关 键 词：芦荟大黄素 胶质母细胞瘤 CASPASE-3 GSDME 焦亡 

分 类 号：R730.5[医药卫生—肿瘤] R285.5[医药卫生—临床医学] R2-031